CN106978367A - A kind of urea Bacillus strain TB42 and its application in compost maturity is promoted - Google Patents
A kind of urea Bacillus strain TB42 and its application in compost maturity is promoted Download PDFInfo
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Abstract
The invention belongs to plant growth-promoting bacteria development technique field, and in particular to be a kind of urea Bacillus strain TB42 and its promote compost maturity in application.A kind of urea Bacillus strain TB42, its taxology is named as urea bacillus TB42 (Ureibacillus suwonensis), China Committee for Culture Collection of Microorganisms's common micro-organisms center was preserved on 01 05th, 2017, preserving number is CGMCC 13529.The urea Bacillus strain TB42 of the present invention is screens isolated from compost megathermal period sample first, the bacterial strain high temperature resistant, increases breeding soon, and can produce lignocellulolytic enzymes；The TB42 of Bacillus strain containing urea microbial inoculum can improve compost temperature, accelerate organic matter degradation and water-soluble organic matter (DOM) degraded, improve the content of kjeldahl nitrogen, promote compost maturity；Also there is the function of biosurfactant production, therefore, the bacterial strain can be used for preparing surfactant.
Description
Technical field
The invention belongs to plant growth-promoting bacteria development technique field, and in particular to be a kind of urea Bacillus strain TB42 and
Its promote compost maturity in application.
Background technology
Chemical fertilizer plays great function in modern agricultural production.But with the increase of fertilizer application amount, its utilization rate
Reduce year by year.The residual of agricultural chemicals and chemical fertilizer, the substantial amounts of harmful substance to Environment release, pollutes soil, water source and food, right
Human health and living environment constitute great threat, it is desirable to preserve the ecological environment increasingly strong with the cry of production safety food
It is strong.Therefore, the new source of manure for substituting chemical fertilizer is developed, to adapt to be to work as the need for developing green agriculture and pollution-free food
Business is anxious.And microbial manure can make invalid nutrition validation in soil, prevention and control corps diseases, agricultural chemicals and change are reduced
The use of fertilizer, is the basic by way of being generally considered a kind of environment-friendly, economy has of solution soil, water source and food pollution
The method of the raising crop yield of effect.
Microbial manure is the particular product that a class contains living microorganisms, applied in agricultural production, results in spy
Fixed fertilizer effect.Wherein viable microbial plays a crucial role in product.At present, that microorganism fertilizer material products typically are divided into two is big
Class：One class is the microbial manure of narrow sense, refers to the vital movement by microorganism, adds the supply of plant nutrient,
Include the total supply of plant nutrient in soil and production environment, cause the improvement of plant nutrient situation, and then increase production
Amount.The representative of this quasi-microorganism fertilizer is rhizobia fertilizer；Another kind of is the microbial manure of broad sense, is referred to by wherein microorganism
Vital movement, can not only improve the supply of plant nutrient, moreover it is possible to produce auxin, promote Plant To Nutrient
The pathogenic effects for absorbing or having some pathogenic microorganisms of antagonism of element, mitigate diseases and pests of agronomic crop brief introduction raising and make produce
Amount.Compared with chemical fertilizer, microbial manure has advantages below：Do not destroy soil texture；It is protecting ecology, free from environmental pollution,
It is nontoxic to people and animals；Fertilizer efficiency is lasting；Crop yield is improved, improves crop quality；It is with low cost, economical and effective.
Plant growth-promoting rhizobacteria (Plant growth promoting rhizobacteria, PGPR) is that a class can be highly dense
Degree colonizes the microorganism in plant rhizosphere, has suppression phytopathogen, rhizosphere harmful microorganism concurrently, and promote plant growth simultaneously
Increase the effect of crop yield.As the valuable source storehouse of bio-feritlizer and biological pesticide, PGPR research and application have risen
Very important effect is arrived.Microbial manure is researched and developed from the angle of resource regeneration then to comprehensive utilization of resources
Realistic meaning is had more with environmental protection.
Bacillus belongs to Bacillaceae, bacillus, can form the Gram-positive bar of endospore-gemma
Bacterium, aerobic or facultative aerobic, with flagellum.Bacillus is distributed widely in soil, empty gas and water and animal intestinal tract, and it is stable
Property it is strong, gemma is just resistant to various unfavorable conditions once being formed, such as xeothermic (150 DEG C of xeothermic 1h still have gemma survival), it is damp and hot,
Ultraviolet, strong acid, highly basic, extremely organic solvent, drying, vacuum drying, oxidation of oxidant etc..Under certain condition,
Gemma can be preserved for a long time, it is ensured that the term of validity of probiotic composition.Bacillus is optimal micro- in all Pseudomonas
Bio-additive.
Lot of experiments proves that bacillus additive can obviously improve flora in body and constitute and with somatotrophic work(
Effect, and bacillus has higher stability in particulate material, the process of powder and in sour environment, in enteron aisle
It can play a role and breed in environment, meet the condition of microbe additive.Bacillus has height in intestinal acidity environment
The stability of degree, can secrete more strongly active protease and amylase, promote the digestion of feed nutrition material.Bacillus can be with
Reduce the ETEC quantity in excrement and alimentary canal.
At present, the Bacillus strain used mainly has bacillus subtilis (Bacillus subtilis), lichens gemma
Bacillus (Bacillus licheniformis), bacillus cereus (Bacillus cereus), Japan bacillus
(Bacillus toyoi) etc., and be then rarely reported on the research of urea bacillus, such as application number 201210222021.9
Patent of invention《Thermophilic urea bacillus and its microbial inoculum and application》One plant of thermophilic urea bacillus JD-50 is disclosed, its preservation is compiled
Number it is CGMCC No.5818, and discloses its application in oil exploitation.
The information for being disclosed in the background section is merely intended to understanding of the increase to the general background of the present invention, without answering
When the prior art for being considered as recognizing or implying the information structure in any form well known to persons skilled in the art.
The content of the invention
Present invention solves the problem in that providing a kind of urea Bacillus strain TB42, the bacterial strain can be applied to prepare surface
Activating agent and promotion compost maturity.
The purpose of the present invention is achieved through the following technical solutions：
The invention discloses a kind of urea Bacillus strain TB42, its taxology is named as urea bacillus TB42
(Ureibacillus suwonensis), China Committee for Culture Collection of Microorganisms was preserved on 01 05th, 2017
Common micro-organisms center, preserving number is CGMCC 13529.
Present invention also offers purposes of the described urea Bacillus strain TB42 in lignocellulose degradation.
Present invention also offers purposes of the described urea Bacillus strain TB42 in surfactant is prepared.
Laccase, lignin peroxidase, manganese are being prepared present invention also offers described urea Bacillus strain TB42
Purposes in peroxidase, cellulase and hemicellulase.
Present invention also offers a kind of microbial inoculum, the urea Bacillus strain TB42 seed liquors described in claim 1 are pressed 1%
(v/v) inoculum concentration, is inoculated into the LB nutrient solutions after 1L sterilizings, and the culture 24h in shaking table (50 DEG C, 120rmp) collects training
Nutrient solution, produces microbial inoculum.
Present invention also offers purposes of the described microbial inoculum in compost maturity is promoted.
Compared with prior art, the present invention has following beneficial technique effect：
1. the urea Bacillus strain TB42 that the present invention is provided is to screen isolated from compost megathermal period sample first
, the bacterial strain high temperature resistant, growth and breeding is fast, and can produce lignocellulolytic enzymes.
2. the microbial inoculum for the TB42 of Bacillus strain containing urea that the present invention is provided can improve compost temperature, accelerate organic matter drop
Solution and DOM degradeds, improve the content of kjeldahl nitrogen, promote compost maturity.
3. the urea Bacillus strain TB42 that the present invention is provided also has the function of biosurfactant production, therefore, should
Bacterial strain can be used for preparing surfactant.
Preservation information explanation
Urea bacillus TB42 (Ureibacillus suwonensis), deposit number is CGMCC 13529, preservation day
Phase is on 01 05th, 2017, and depositary institution is China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation
Address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica.
Brief description of the drawings
Fig. 1 is temperature with compost time changing curve.RT：Room temperature, blank group (CK), experimental group (T)
Fig. 2 is pH with compost time changing curve.Blank group (CK), experimental group (T)
Fig. 3 is moisture with compost time changing curve.Blank group (CK), experimental group (T)
Fig. 4 is the content of organic matter with compost time changing curve.Blank group (CK), experimental group (T)
Fig. 5 is kelvin nitrogen content with compost time changing curve.Blank group (CK), experimental group (T)
Fig. 6 is ammonia-nitrogen content with compost time changing curve.Blank group (CK), experimental group (T)
Fig. 7 is nitrate nitrogen content with compost time changing curve.Blank group (CK), experimental group (T)
Fig. 8 is DOM contents with compost time changing curve.Blank group (CK), experimental group (T)
Fig. 9 is Ureibacillus suwonensis TB42 bacterium colony figures.
Figure 10 is performances of the Ureibacillus suwonensis TB42 on blood agar plate.
Figure 11 is glucose Standard for Sugars curve.
Figure 12 is xylose standard curve.
Figure 13 is the gel electrophoresis spectrum for filtering out bacterial strain；
About the explanation of reference：
M-2000bpMake；1-TB21；2-TB22；3-TB23；4-TB24；5-TB25；6-TB41；7-TB42；8-TB43；
9-TB44；10-TB45；11-TB61；12-TB62；13-TB63；14-TB64；15-TB65；16-TA65；17-TB46.
Embodiment
With reference to specific embodiment, the present invention is described in further detail, it is described be explanation of the invention and
It is not to limit.
Material and reagent used, unless otherwise specified, are commercially obtained in following embodiments.It is following to implement
Experimental method used in example is conventional method unless otherwise specified.
Embodiment 1：Urea bacillus TB42 (Ureibacillus suwonensis) screening
1. experiment material and equipment
1.1 main agents
The main agents of table 1
1.2 key instruments and equipment
The key instrument of table 2 and equipment
1.3 culture medium
Bacteria culture media：Nutrient agar 33g, deionized water 1000mL；
LB nutrient solutions：Peptone 10g, dusty yeast 5g, sodium chloride 10g, deionized water 1000mL.
2. the separation screening of lignin-degrading bacteria
2.1 sampling
The sample of sieve bacterium is respectively 2, the compost sample of 4,6 days, in heap temperature higher position multidraw (typically in heap body
Lower face 10-20cm), it is fitted into sample sack after sample blending in being preserved in -4 DEG C of refrigerator.
The screening and purifying of 2.2 high temperature resistant bacterium
Sample 10g is weighed, is added in the triangular flask containing the NaCl solution (0.9%w/v) after 90mL sterilization treatments, puts
In vibrating 30min on shaking table, it is therefore an objective to break up zoogloea, bacterium is scattered in unicellular in solution.By sterilization treatment
NaCl solution afterwards is added separately in 7 test tubes (same sterilization treatment is crossed), every addition 9mL, takes 1mL points of bacterium solution after concussion
Test tube is not added, i.e., be diluted to 10 respectively-1、10-2、10-3、10-4、10-5、10-6、10-7, then respectively by the bacterium solution under each dilution factor
0.1mL is taken to be coated on bacteria culture media flat board, 50 DEG C incubated.The flat board of every kind of dilution gradient make 5 it is parallel.Can be with
It was observed that bacterial community is 10-5Growing way is preferable under dilution factor.The larger carry out separation training of bacterium colony is chosen in five parallel flats
Support and repeatedly line purifying.
As shown in figs. 9-10, urea bacillus TB42 colonial morphologies are irregular, and bacterium colony is larger, the smooth moistening in surface, semi-transparent
Bright, milky, colony edge is irregular.
3. crude enzyme liquid is produced
LB nutrient solutions are prepared, sterilization treatment is poured into same sterilized serum bottle (25mL), isolated strain bacterium
It is inoculated into serum bottle, is put into shaking table (50 DEG C, 120rmp) culture, prepares seed liquor.LB nutrient solutions are prepared, are fallen in conical flask
Enter 100mL, sterilization treatment draws above-mentioned 1mL seed liquors and shaking table culture (50 DEG C, 120rmp) is put into conical flask, equally, makes
Into bacterium solution.
Liquid producing enzyme culture：Bacterium solution centrifuges 5min through 5000r/mim, supernatant is taken as crude enzyme liquid, with heat inactivation enzyme liquid
(taking 0.6mL crude enzyme liquids in centrifuge tube, to boil 10min) is used as control.
4. the assay method of enzyme activity
The measure of 4.1 laccases (Lac) activity
1st, reagent
(1) determined by oxydasis ABTS speed；
(2) 0.1moL/L citric acid-sodium citrate buffer solutions:2.941g sodium citrates and 2.1014g lemons are weighed respectively
Acid, plus deionized water 100mL, the adding citric acid into sodium citrate adjust pH=5.0 with pH meter；
(3)0.5mL 0.5moL/L ABTS；
2nd, operating procedure
(1) at 25 DEG C, 2mL citric acid-sodium citrate buffers (0.1mmoL/L, pH=5) are added in test tube,
With 0.5mL ABTS, mixed liquor is poured into cuvette；
(2) extinction value changes and then addition 1mL enzyme liquids start reaction, are determined under 420nm.
3rd, computational methods
Enzyme activity is defined：ABTS for 1 μm of oL of oxidation per minute is an enzyme-activity unit,
Extinction coefficient epsilon=3.6*104[(moL/L)-1cm-1], enzyme-activity unit：U/L
Laccase (Lac) vigor=Δ OD6/ΔT*VEnzyme liquid*∈
=(Δ OD/ Δ T) * 106/0.5*3.6*104
In formula：The value for the adjacent absorbance value difference maximum that Δ OD- is determined；The corresponding time difference of Δ T- Δ OD values；
The volume of V- enzyme liquids.
The measure of 4.2 lignin peroxidases (Lip) activity
1st, reagent
(1) 0.24moL/L sodium tartrates buffer solution：3.60216g tartaric acid and 5.52192g sodium tartrates are weighed respectively,
Plus deionized water 100mL, tartaric acid is added into sodium tartrate, pH=3.0 is adjusted；
(2) 0.1mL 24mmoL/L veratryl alcohols；
(3) 0.05mL 6.0mmoL/L H2O2；
2nd, operating procedure
(1) at 37 DEG C, overall reaction system is 3mL, and 1.85mL 0.24moL/L sodium tartrates buffering is added into test tube
Liquid (pH=3.0), and 0.1mL 24mmoL/L veratryl alcohols and 1.0mL enzyme liquids；
(2) 37 DEG C are preheated to solution is poured into cuvette, 0.05mL 6.0mmoL/L H is then added again2O2Start anti-
Should, determine extinction value changes in the case where wavelength is 310nm.
3rd, computational methods
It is an enzyme-activity unit that enzyme activity, which is defined as 1 μm of oL of oxidation per minute veratryl alcohol,
Extinction coefficient epsilon=9.3*103[(moL/L)-1cm-1].Enzyme-activity unit：U/L
Peroxidase (Lip) vigor=Δ OD*106/ΔT*VEnzyme liquid*∈
=(Δ OD/ Δ T) * 106/1*9.3*103
In formula：The value for the adjacent absorbance value difference maximum that Δ OD- is determined；The corresponding time difference of Δ T- Δ OD values；
The volume of V- enzyme liquids.
The measure of 4.3 manganese peroxidases (Mnp) activity
1st, reagent
(1) 0.11moL/L sodium lactate buffer solution：1.10098g lactic acid and 2.05516g sodium lactates are weighed, adds go respectively
Ionized water 100mL, lactic acid is added into sodium lactate, adjusts PH=4.5；
(2) 0.025M l40mmoL/L manganese sulfates；
(3) 0.025mL 1.6mmoL/L H2O2；
2nd, operating procedure
(1) according to enzyme in H2O2In the presence of Mn2+Aoxidize Mn3+Speed determine；
(2) total reaction volume is 1mL, and 0.85mL 0.11moL/L sodium lactate buffer solution (pH=is added into test tube
4.5) 0.025mL, 40mmoL/L MnSO are added4With 1mL enzyme liquid；
(3) preheat after 37 DEG C, solution is poured into cuvette, 0.025mL 1.6mmoL/L H is added in cuvette2O2Open
Dynamic reaction, determines extinction value changes under 240nm.
3rd, computational methods
The definition of enzyme activity is 1 μm of oL of oxidation per minute Mn2+For Mn3+For an enzyme-activity unit,
Extinction coefficient epsilon=6.5*103[(moL/L)-1cm-1].Enzyme-activity unit：U/L
Manganese peroxidase (Mnp) vigor=Δ OD*106/ΔT*VEnzyme liquid*∈
=(Δ OD/ Δ T) * 106/1*6.5*103
In formula：The value for the adjacent absorbance value difference maximum that Δ OD- is determined；The corresponding time difference of Δ T- Δ OD values；
The volume of V- enzyme liquids.
The measure of 4.4 cellulase activities
1st, reagent
(1) 0.1moL/L sodium citrate-citric acids sodium buffer solution：2.941g sodium citrates and 2.1014g lemons are weighed respectively
Lemon acid, plus deionized water 100mL, the adding citric acid into sodium citrate adjust pH=4.8 with pH meter；
(2)1.5mLDNS；
(3) glucose standard：The deionized water that 1.0000g glucose is dissolved in 1000ml is configured to 1mg/mL glucose
Titer.
2nd, standard glucose curve plotting
1mg/mL standard glucose liquid glucoses each 0 are taken, 0.2,0.4,0.6,0.8,1.0,1.2mL in test tube, adds deionization
Water adds 2.0mL DNS reagents to 2.0mL, and tool plug, boiling water bath 10min is settled to 15mL after cooling, uses spectrophotometer
OD values are surveyed in the case where wavelength is 550nm, 3 repetitions are tested, and take average to chart, glucose Standard for Sugars curve is referring to Figure 11.By optical density
Value invites glucose amount.
3rd, operating procedure
(1) 1cm*2cm Whatman NO1 quantitative test papers are added into test tube, 1.0cm sodium citrates-lemon is added
Sour sodium buffer solution (0.1moL/L, pH=4.8) and 0.5mL enzyme liquids；
(2) 1h is incubated at 50 DEG C, then takes out flowing water after addition 1.5mL DNS terminating reactions, then waste water bath 5min cold
But, it is settled to 25mL；
(3) its absorbance is determined under 540nm.
4th, computational methods
Cellulase activity=y*1000ug/VEnzyme liquid/ T, wherein y obtain y=0.6916x by mark song
In formula：The absorbance that X- is determined；The time of T- water-baths；VEnzyme liquidThe volume of-enzyme liquid.
4.5 hemicellulase activities are determined
1st, reagent
(1) 1.8mL 1% xylan solution：1% solution is made into pH=4.8 acetate buffer solutions with xylan；
(2) 1.8mL acetate buffer solutions：0.82g sodium acetates are weighed, 0.6mL acetic acid is measured, deionized water 100mL is added respectively,
Add acetic acid into sodium acetate, adjust pH=4.8；
(3)2mL DNS；
(4) Xylose Standard：The deionized water that 1.0000g xyloses are dissolved in 1000mL is configured to 1mg/mL xylose standard
Liquid.
2nd, xylose curve plotting
Take 1mg/ml standard Xyloses each 0,0.2,0.4,0.6,0.8,1.0,1.2mL in test tube, add deionized water
To 2.0mL, 2.0mL DNS reagents are added, tool plug, boiling water bath 10min are settled to 15mL after cooling, existed with spectrophotometer
Wavelength is survey OD values under 550nm, and 3 repetitions are tested, and take average to chart, xylose standard curve is referring to Figure 12.Drawn by OD value
Obtain xylose amount.
3rd, operating procedure
(1) enzyme liquid 0.2mL, then each xylan solution for drawing 1.8mL 1% are added in 15mL scale test tubes；
(2) blank adds 1.8mL acetate buffer solutions with 0.2mL enzyme liquids, is not added with xylan solution, shakes up；
(3) 50 DEG C of water-bath 60min, after taking-up, then draw 2mL DNS reagents and shake up, tool plug, and boiling water bath reacts immediately
10min, after cooling, adds water constant volume to 15mL, gently shakes up up and down；
(4) blank zeroising is used, in surveying absorbance under wavelength 550nm.
4th, computational methods
Calculated according to formula：Hemicellulose enzyme activity=y*1000ug/VEnzyme liquid/ T, wherein y obtain y=1.1504x by mark song
+0.008；In formula：The absorbance that X- is determined；The time of T- water-baths；VEnzyme liquidThe volume of-enzyme liquid.
5th, result
The Ureibacillus suwonensis TB42 strain enzyme-producing determinations of activity of table 3
As shown in table 3, bacillus licheniformis TB42 produces the vigor highest of manganese peroxidase, is 207U/L, next to that fine
The plain enzyme of dimension, its vigor is 78U/L, and the vigor of laccase, lignin peroxidase and hemicellulase is respectively 7U/L, 0U/L
And 16U/L.
Embodiment 2：Bacillus licheniformis TB42 (Ureibacillus suwonensis) identification
2.1 experiment key instruments
Table 4 tests instrument
2.2 experimental procedure
2.2.1 DNA is extracted
DNA uses raw work《Ezup pillar bacterial genomes DNA extraction agent box specifications》Extract.Concrete operation step is detailed
See specification.
2.2.2 PCR is expanded
Enter performing PCR using 2 × Es Taq MasterMix to expand, reaction system is as follows：
Response procedures：
2.2.3 PCR primer electrophoresis result
Using the agarose gel electrophoresis of 1% concentration, voltage 120V, electrophoresis 30min, per hole loading 1uL, electrophoresis pattern is shown in
Figure 13.
As shown in figure 13, PCR primer electrophoresis result shows the 16S rDNA fragments of Successful amplification bacterium.
2.2.4 sequencing and comparison result
Sequencing uses the two-way sequencings of primer 2 7F/1492R, and urea Bacillus strain TB42 sequence is as shown in SEQ.1.
BLASTN comparisons are carried out in NCBI gene pools, strain idenfication result is obtained, the results are shown in Table 5.
The urea Bacillus strain TB42 appraising datums of table 5
As shown in Table 5, bacterial strain TB42 reaches 99% with Ureibacillus suwonensis similarities, therefore, by bacterial strain
It is named as urea bacillus TB42 (Ureibacillus suwonensis).
Embodiment 3：Application of the TB42 of Bacillus strain containing the urea microbial inoculum in compost maturity is promoted
It is prepared by 3.1 microbial inoculums
TB42 seed liquors are pressed 1% (v/v) inoculum concentration, be inoculated into the LB nutrient solutions after 1L sterilizings, in shaking table (50
DEG C, 120rmp) middle culture 24h, it is microbial inoculum to collect nutrient solution.It is experimental group (T), blank group (CK) addition 1L to add microbial inoculum group
LB nutrient solutions after sterilizing.
3.2 compost are tested
Composting material selects cow dung and sugarcane top, mass ratio 17:3, common 20kg.Compost carries out 45d, respectively the 0th, 5,
10,16,23,30,45d samplings, determine temperature, pH, moisture content, organic matter, kjeldahl nitrogen, inorganic nitrogen, the parameter such as DOM contents.
Whole composting process is inoculated with microbial inoculum twice, respectively in 0d and 10d.Heap body turning three times, is 10d, 20d and 30d respectively.
3.3 experimental result
Urea bacillus TB42 high temperature resistants, increase breeding soon, and can produce lignocellulolytic enzymes.
Compared with the compost without microbial inoculum (CK), the compost (T) of addition TB42 microbial inoculums has the advantage that：
(1) in terms of the temperature of composting process, the heap temperature of addition TB42 microbial inoculums compares the heap temperature without microbial inoculum
Significantly improve.This is conducive to killing the high temperature degradation of pathogen and acceleration compost material in heap body, promotes heap body to become thoroughly decomposed (Fig. 1).
(2) by Fig. 2, Fig. 3 is understood, the pH for adding the compost of TB42 microbial inoculums rises get Geng Gao, and moisture drops faster, that is, adds bacterium
The heap precursor reactant of agent is more acutely so as to cause NH3A large amount of volatilization increase pH, heat production is more to take away substantial amounts of moisture.
(3) from the point of view of heap body dissolved organic matter (DOM) changes of contents, without microbial inoculum DOM contents by
69.6mg/g drops to 9.8mg/g, have dropped 85.9%, the DOM contents of addition TB42 microbial inoculums are dropped to by 68.8mg/g
6.7mg/g, have dropped 90.3% (Fig. 8).
In summary, addition TB42 microbial inoculums can improve compost temperature, accelerate moisture loss, accelerate organic matter degradation and
DOM degrades, and promotes compost maturity.
Embodiment 4：Urea Bacillus strain TB42 produces surfactant situation
By blood agar plate and surface tension instrument, three plants of bacterium of identification produce surfactant situations jointly.
Blood agar plate preparation method：Take after nutrient agar (Beijing overpass), sterilization treatment, treat that it is cooled to 50 DEG C or so,
Gnotobasis adds the de- fiber Sheep Blood 5-10mL of sterilizing in every 100mL nutrient agars, gently shakes up, flat board is poured at once, make
It is standby into inclined-plane.
Zymotic fluid surface tension test：Using around-France measure zymotic fluid surface tension, using full-automatic surface tension instrument, instrument
Type number：BZY-1.
It is demonstrated experimentally that urea Bacillus strain TB42 can produce surfactant.
It is foregoing to the present invention specific illustrative embodiment description be in order to illustrate and illustration purpose.These descriptions
It is not wishing to limit the invention to disclosed precise forms, and it will be apparent that according to above-mentioned teaching, can be much changed
And change.The purpose of selecting and describing the exemplary embodiment is that explaining that the certain principles and its reality of the present invention should
With so that those skilled in the art can realize and using the present invention a variety of exemplaries and
A variety of selections and change.The scope of the present invention is intended to be limited by claims and its equivalents.
SEQUENCE LISTING
<110>Guangxi University
<120>A kind of urea Bacillus strain TB42 and its application in compost maturity is promoted
<130> ZYWS
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1458
<212> DNA
<213>Artificial sequence
<400> 1
cgggggggga tgcctataca tgcaagtcga gcggaccaat tagaaagctt gctttttaat 60
tggttagcgg cggacgggtg agtaacacgt gggtaacctg ccctatagac cgggataact 120
cgcggaaacg cgtgctaata ccggataaca caccgaagcg catgcttcgg ggttgaaaga 180
tggttctgct atcactatag gatgggcccg cggcgcatta gctggttggt ggggtaacgg 240
cctaccaagg cgacgatgcg tagccgacct gagagggtga tcggccacac tgggactgag 300
acacggccca gactcctacg ggaggcagca gtagggaatc ttccacaatg ggcgaaagcc 360
tgatggagca acgccgcgtg agcgaagaag gtcttcggat cgtaaagctc tgttgtaagg 420
gaagaacaag cgcagcagtc actggctgcg ccctgacggt accttactag aaagccacgg 480
ctaactacgt gccagcagcc gcggtaatac gtaggtggca agcgttgtcc ggaattattg 540
ggcgtaaagc gcgcgcaggc ggtctcttaa gtctgatgtg aaagcccccg gctcaaccgg 600
ggagggtcat tggaaactgg gagacttgag tgcaggagag ggaagyggaa ttccatgtgt 660
agcggtgaaa tgcgtagaga tatggaggaa caccagtggc gaaggcggct tcctggcctg 720
taactgacgc tgaggcgcga aagcgtgggg agcaaacagg attagatacc ctggtagtcc 780
acgccgtaaa cgatgagtgc taggtgttag ggggtttccg ccccttagtg ctgcagctaa 840
cgcattaagc actccgcctg gggagtacgg tcgcaagact gaaactcaaa ggaattgacg 900
ggggcccgca caagcggtgg agcatgtggt ttaattcgaa gcaacgcgaa gaaccttacc 960
aggtcttgac atcccgctga ccgccatgga gacatggctt tcccttcggg gacagcggtg 1020
acaggtggtg catggttgtc gtcagctcgt gtcgtgagat gttgggttaa gtcccgcaac 1080
gagcgcaacc cttgtcctta gttgccatca ttcagttggg cactctaagg agactgccgt 1140
acaaatacgg aggaaggtgg ggatgacgtc aaatcatcat gccccttatg acctgggcta 1200
cacacgtgct acaatgggtg gtacaaaggg cggcaaaccc gcgaggggga gcgaatccca 1260
aaaagccact ctcagttcgg attgcaggct gcaactcgcc tgcatgaagc cggaatcgct 1320
agtaatcgcg gatcagcatg ccgcggtgaa tacgttcccg ggccttgtac acaccgcccg 1380
tcacaccacg agagtctgta acacccgaag tcggtgaggt aaccctccgg gagccagccg 1440
ccgaaaggtg acccgagt 1458
Claims (6)
1. a kind of urea Bacillus strain TB42, its taxology is named as urea bacillus TB42 (Ureibacillus
Suwonensis), China Committee for Culture Collection of Microorganisms's common micro-organisms center was preserved on 01 05th, 2017,
Preserving number is CGMCC 13529.
2. purposes of the urea Bacillus strain TB42 according to claim 1 in lignocellulose degradation.
3. purposes of the urea Bacillus strain TB42 according to claim 1 in surfactant is prepared.
4. urea Bacillus strain TB42 according to claim 1 is preparing laccase, lignin peroxidase, manganese peroxide
Purposes in compound enzyme, cellulase and hemicellulase.
5. a kind of microbial inoculum, it is characterised in that：Urea Bacillus strain TB42 seed liquors described in claim 1 are pressed into 1% (v/v)
Inoculum concentration, be inoculated into the LB nutrient solutions after 1L sterilizings, the culture 24h in the shaking table (50 DEG C, 120rmp) collects nutrient solution,
Produce microbial inoculum.
6. purposes of the microbial inoculum according to claim 5 in compost maturity is promoted.
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| CN113773987A (en) * | 2021-08-26 | 2021-12-10 | 郑州轻工业大学 | Biological agent for improving aerobic fermentation efficiency of organic waste and preparation method thereof |
| CN114380629A (en) * | 2022-02-17 | 2022-04-22 | 重庆市农业科学院 | Composite microbial straw decomposing inoculant and preparation method thereof |
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